DEVELOPMENT OF AN ETHANOL PRODUCTION PROCESS WITH STILLAGE

RECYCLING AT PILOT PLANT-SCALE.

Castro, G. A.a, Gil, I. D.b

a
Universidad Nacional Abierta y a Distancia, Escuela de Ciencias Básicas, Tecnología e
Ingeniería Calle 14 sur Nº 14-23, Bogotá, Colombia.
b
Department of Chemical Engineering, Universidad de los Andes , Calle 19A No. 1-37 Este,
Bogotá, Colombia

Abstract

The production of energy from biomass has been object of wide research in the last years,
especially the production of fuel alcohol, in Colombia, has increased thanks to laws
promulgated by the government; parallel to this the generation of residues increases also, in
this case called stillage or vinasse, for a conventional process 13 to 15 liters are obtained of
stillage by liter of ethanol, which for the quantity of ethanol projected, of 750 million liters/
year of alcohol in this country, would be generated between 9750 and 11250 million liters of
stillage/year, a so big quantity. An ethanol production process with stillage recycling in
bench-scale was developed in order to determine stillage percentage recirculation and the
number of successive recycles that can be carried out, without detriment in the ethanol
productivity achieved. With the best results obtained in bench-scale will be climbed to pilot
plant-scale. The inhibitors presents in the stillage, as acetic acid and solids, which increase by
accumulation, with the continuous recycles, limit the recirculate percentage and the number
of recycles that can be carried out, for this reason several alternatives were proposed and
evaluated in order to remove them or diminish their harmful effect.

Keywords: stillage recycling, ethanol production, clean technology, pilot plant scale.

1. Introduction

The great demand of alcohol that actually we can observe causes, as response of the industry,
a high production with the subsequent generation of residues as the vinasse or stillage, to
which it is necessary to give her the importance that is deserved, as pollutant potential or
source of by-products if this way it is wished.

As answer to the world critical environmental situation, to the gradual decrease of the fossil
fuels and the possibility in the country of developing alternative energies using renewable
resources, the government determined to send the Law 693 of 2001, where is regulated the
addition of anhydrous ethanol to the gasolines in 10% starting from 2006, according to this it
is necessary to increase the production of this alcohol, until levels initials of 750 million liters
/ year, (Program of Carburating Alcohols).
As agricultural sources for the obtaining of the ethanol there are the sugary ones (cane of
sugar, beet), the starch sources (barley, banana tree, yucca), the lignocellulosics (wood,
cotton). The physicochemical characteristic of all sources are different whose causes
differences in the adaptation operations for a common later treatment in the three cases,
whichever it is the source of sugar for the fermentation, the processes have the high
production of liquid effluents of the distillation in common, denominated in this particular
case, vinasses or stillage, Durán de Bazúa in 1993 defined the vinasse like "a dilution of
substances like mineral and organic salts with relative value and with potential for diverse
uses", thanks to his organic high load (6.5.7.6%), CaO (4.57%) and K2O (1.06-1.53%)
(GEPLACEA / PNUD / ICIDCA 1990), among other substances, that make it appropriate of
being used as energy source, protein source, mineral salts source for fertilizers and
concentrate for animals.

In the conventional process of production of alcohol the vinasse production oscillates

between 13 and 15 l/l ethanol, with this and if there are a requirement of 2 million liters of
ethanol / day approximately, according to the Carburating Program of Alcohol, the quantity
of the effluent per day could ascend to 30 million liters, that it is an important quantity and
that could harm a lot in the environment, if it isn’t managed appropriately like some
evidences show us for example the experiences of the Cauca`s river Valley, that which can
provoke the cancellation of the Environmental License in these distilleries.

As alternative to reduce the vinasse volume has been propossed the recirculation (recycling)
from the same ones to the fermentation stage, like dilution water, however the presence of
inhibitors like the acetic acid, limits their application. The present work evaluates the effect
of the recirculation on the alcoholic yield and biomass generation in the fermentation.

2. Materials and Methods

2.1 Culture
The used culture was a Saccharomyces cerevisiae ATCC26603, donated by the Institute of
Biotechnology of the National University of Colombia (IBUN), in liofilized state, it was
resuspended in peptoned water
and it was sowed in a growth medium (yeast extract 1%, peptone 2%, glucose 1%, maltose
1%, agar 2%, water 93%, all percentages w/w) at 30ºC for 48 h, starting from this cultivation
broth was prepared the main cultivation (peptone 1%, sucrose 2%, K2HPO4 0.2%, MgSO4
0.001%, (NH4)2SO4 0.2%, water 96.5%, pH 4.5) and was inoculated with 15% of the
previous broth and it was renewed monthly.

2.2 Fermentation and distillation

The inoculum (4.5 L)for the fermentation, constituted 15% of the broth of fermentation(30
L), both were compound of cane molasses 16.5%, water 83.5%, (NH4)2SO4 0.01%, the pH
was adjusted at 4.5, with a solution 15% w/w of H2SO4, the time of preparation of the
inoculum was of 14 h and that of the fermentation of 28 h in a CE 640 Fermentation &
Alcohol Production Pilot Plant / PC (Gunt, Hamburg) with temperature control at 30ºC and
110 rpm, after the fermentation time, the fermented must was pumped to the distillation unit,
there was carry out the thermal process during 6 hours with a vapor pressure of 250 kPa.
2.3 Vinasse treatment
The vinasse remains in the boiler after the distillation, then it’s carry to a continuous
centrifuge at 1200 g, the light phase in then ready to be recycled, replacing 70% of the
dilution wáter.

2.7 Analysis
The kinetics of the fermentation was verified by analysis of sugars and etanol at different
fermentation times by high performance liquid chromatography (HPLC) in a Waters
chromatography system, with a detector of refraction index, each sample was centrifuged 900
g during 15 minutes, later it was diluted ten times, starting from the second fermentation a
solid phase extraction was applied using Sep pack C18, to retire the extractant agent's
residuals that could remain in the recycled vinasse, after the sample was filtered using a 0.22
µm membrane and finally it was injected manually in the cromatograph. The used column
was a Sugar-Pack type Waters, as mobile phase it was used water deionized and degasifed
with a flow of 0.5 ml / min, the column temperatures and detector were respectively of 84
and 40ºC, the volume of sample was 20 µl.

The biomass was determined, using the technique of the camera of Neubawer.

3. Results and Analysis

The fermentative capacity of the yeast was evaluated previously (Castro, 2006) and in this
work was observed the soluble solids decrease with the fermentation time in the CE 640
Fermentation & Alcohol Production Pilot Plant / PC (Gunt, Hamburg), the results are
observed in the figure 1.

Figure 1 Soluble solid decrease with the fermentation time

 Figure 2 Ethanol final concentration of the fermented musts prepared with recycled vinasse

The figure 2 shows how varies the concentration of ethanol in each one of the five
fermentations in series, with recirculation of 70% of the vinasse only applying the
centrifugation, is posible to see a similar behavior in the work of Castro et. al. at 2006
but with a smaller final etanol concentration, of 36.8 g/L front 40.2% of the previous
work.

The figure 3 shows something similar to the figure 2 to the ethanol, this time with the
biomass, in this case it presents a continuous descent, making notorious that the answer
of the yeast is more sensitive than that of production of etanol, in the first case the
reduction wasof 80%, in thesecond only of 40%. The results are very close to that
obtained by Castro(2006),

Figure 3 Final biomass in the fermented musts prepared with recycled vinasse.

Besides the inhibitory effect for the fermentation by-products, is very important the
effect of the increase of the solids in the vinasse, they can includ the not fermentable
sugars that remain in the vinasse after the fermentation and the distillation, mineral salts
not assimilated by the yeast, are also included, these substances are increased with the
advance of the serial fermentations, due to the molasses and the nutrients with those the
mediums of fermentation are prepared are rich in these substances (Maiorella, 1984),
others are by-products of the metabolism of the yeast like glicerol, propanol, furfural
and lactic acid that also inhibit the fermentation and the speed of growth (Navarro, 1989
and Maiorella 1983).

Figure 4 Increase in the soluble solids in the médiums prepared with recycled vinasse

In the figure 4 the increase of solids is observed in the mediums that were prepared with
the recycled vinasse, t7he soluble solids (fermentable coming from the molasses and not
fermentable coming from the vinasses) in the mediums of the fifth fermentation,
reached values of 25ºBx, very close at the toxicity level, of 26% of dry matter presented
for Navarro (2000), knowing that of the total of dry matter in the vinasse the 85 to 90%
constitute the soluble solids, being this another of the possible reasons for the decrease
in the productivity of ethanol and generation of biomass in the fifth fermentation and
still for the fourth, where the values of dry matter are from the order of 23ºBx that are
very near to the toxicity limit value, provoking the same effect on the two mentioned
answer variables. Thatipamala and collaborators in 1992, reached a similar conclusion,
since they verified a decrease of 33% (w/w) in the production of ethanol, when the
content of solids in the fermentation medium increases from 14% to 27% (w/w).

4. Conclusions

The results obtained in laboratory scale are very similar to that in the pilot plant scale.

The fermentation time can be from 20 to 30 hours depending on the scale used.

The ethanol concentration decrease 40% and the biomass 80% with the the five
fermentation in series, the principal reason could be the increase in the soluble solids, in
this case of 40% aproximately.

During the five fermentations it was achieved a saving of water of 48% and a decrease
of the volume of vinasses of 40%.
5. References

1) Bilford H., Scalf R., Stark W., Kolachov P. ; Alcoholic Fermentation of Molasses; Industrial and
Engineering Chemistry Research, Vol. 334, Issue 11, 1942
2) Castro, G..A., Caicedo, L. A., Reduction of stillage’s volumen in the production of etanol by
recirculation applying liquid-liquid extraction, Magister Tesis, National University of Colombia,
2006.
3) Durán de Bazúa, C. 1993. "Tratamiento biológico de aguas residuales de la Industria de proceso".
Facultad de Química. Ciudad Universitaria, 127 pags, México, D.F.
4) Federación Nacional de Biocombustibles, abc de los alcoholes carburantes en la página web:
http://www.minminas.gov.co/minminas/sectores.nsf/2a84e89f4d73f130052567be0052c75a/ffdff1b2
98ac766a052571720071cfad?OpenDocument
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7) Navarro, A.; Sepúlveda, M; Rubio, M.; Bio-concentration of vinasse from alcoholic fermentation of
sugar cane molasses; Waste Management, Vol. 20 pg. 581 – 585, 2000.
8) Obregon, J; Romero, O; Sebrango, C; Curbelo,I; Producción industrial de bioetanol y biomasa de
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Universitario de Sancti Spíritus “ José Martí Pérez”, Cuba 2006.
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Carburantes, Jorge Cárdenas Gutiérrez, Medellín, Junio 10, 2004.
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Kinetics and Biomass and Product Yields during Ethanol Batch Fermentation; Biotechnology and
Bioengineering Vol 40, pg. 289-297, 1992.